Clerico Eugenia M, Ditty Jayna L, Golden Susan S
Department of Biology, Texas A&M University, College Station, USA.
Methods Mol Biol. 2007;362:155-71. doi: 10.1007/978-1-59745-257-1_11.
Synechococcus elongatus PCC 7942 is an excellent model system for studying the molecular mechanism of the circadian clock in cyanobacteria. The "plastic" genetic characteristics of this organism have facilitated the development of various methods for mutagenesis of its chromosome. These methods are based on homologous recombination between the chromosome and foreign DNA, introduced to the cyanobacteria by either transformation or conjugation. Here we describe different approaches to mutagenize the chromosome of S. elongatus, including insertional mutagenesis, hit-and-run allele replacement, rps12-mediated gene replacement, and regulated expression of genes from ectopic sites, the neutral sites of the S. elongatus genome.
聚球藻PCC 7942是研究蓝藻生物钟分子机制的优秀模型系统。该生物体的“可塑性”遗传特性促进了多种染色体诱变方法的发展。这些方法基于染色体与外源DNA之间的同源重组，通过转化或接合作用导入蓝藻。在此，我们描述了诱变聚球藻染色体的不同方法，包括插入诱变、打了就跑等位基因替换、rps12介导的基因替换，以及从聚球藻基因组的中性位点异位位点调控基因表达。
